Treatment of psoriasis



Nov. 23, 1937. M, MARCUS 2,099,696

TREATMENT OF PSQRIASIS iled Oct. 10' 1934 Patented Nov. 23, 1937 UNITEDSTATES PATENT OFFICE 12 Claims.

My invention relates to a method for producing a vaccine for treatingpsoriasis, and to an improved vaccine for treating the said disease.

This application is a continuation in part of my application Serial No.181,974 filed on April One of the objects of my invention is to devise amethod whereby the spores of the micro-organism which produce thedisease can be emciently isolated and propagated to produce a pureculture from which a vaccine can be made for treating the said disease.

Another object of my invention is to provide a method whereby thegerminated spores having the ordinary form of filamentedmicro-organism's can be eificiently isolated to produce a vaccine forthe above mentioned purpose.

Another object of my invention is to provide a vaccine for treating theabove mentioned dis-.

ease, it being understood that the claims for the vaccine per se are notlimited to any particular method of producing the said vaccine.

Other objects of my invention will be set forth in the followingdescription and drawing which illustrates a preferred embodimentthereof, it being understood that the above general statement 7 of theobjects of my invention is intended merely to generally explain thesame, and not to limit it in any manner.

The drawing shows a microphotograph of a mono-spore-culture containingthe spores I and fiamentous micro-organisms 2, and all the otherimportant phases indicated by Nos. 3, 4, 5, 6, 7, 8 and 9. This drawingis of the micro-organism enlarged over 400 diameters.

Psoriasis is a skin disease that has been well recognized and has beenwidespread for a great many years. It manifests itself by the appearanceof papules or patches on any part of the skin, the said patches havingcharacteristic white silvery scales. Up to the present time, a greatmany theories have been advanced to ex plain the cause of this disease,but no definite and clearly proved theory has heretofore beenpropounded, and no definite cure has heretofore been proposed.

I have discovered that the cause of psoriasis is a filamentousmicroorganism which reproduces by means of spores and which has certainwell defined characteristics which will be later explained. I have alsodiscovered that by recovering this in pure culture it can be utilizedfor trea'ment in two ways: (A) by direct inoculation of the livingmicro-organism in tl to form of solid culture, and (B) by injection ofthe non-living micro-organism in vaccine form. Thus both the inoculationon the scarified skin and injections of the produced vaccine have adirect and powerful curative eflect.

Hence, whenever I specify the use of vaccine 5 eitherin the descriptionor in the claims herein, it is understood that I also wish to coverbroadly any inoculation with the living micro-organism.

In order to isolate the filamentous micro-organism which producespsoriasis, the following 10 procedure is effective;-

A lesion on the skin of a patient is first washed with alcohol, followedwith a wash of and then alcohol to remove or destroy the saprophytes andto thoroughly cleanse the lesion. 15 Some of the surface scales are thenimmediately transferred to a suitable culture medium such as ordinaryagar, glucose agar, maltose agar, Sabourauds agar, and other ordinarysugar media. Other surface scales are kept for an hour 20 in 60% alcoholand are then inoculated upon the culture medium.

The second batch of scales which are kept for an hour in 60% alcohol areinoculated upon a batch of culture medium which is different from 25 thefirst batch utilized.

While I wish to have my invention cover the use of scales taken fromsurface lesions, I have discovered that the use of such scales is notnecessary and I do not wish to be limited to the use 30 of such surfacescales.

After the surface scales have been removed from a lesion so that thesaid lesion is substantially free from surface scales, the second levelthus secured is again washed with 70% alcohol 35 and then with 95%alcohol and the skin at the second level is then scarified to cause theoozing of serum or lymph. Enough of this serum is taken to inoculate athird batch of culture media and to prepare impression smears both ofthe 40 serum itself and smears of the mixed serum and the scales takenfrom the first level.

The exposed surface, which now corresponds to what may be called thethird level, is again washed with alcohol as above mentioned; the ex 45posed surface is again irritated; and serum cultures and smears areprepared as above specified in the second stage of my Process.

In order to produce the irritation above mentioned, I prefer to use asteel scalpel having a 50 handle about 5 inches long and a point about 2inches long, one surface of the blade being flat. This scalpel should bekept in a sterilizing solution comprising 95% of phenol and when it isused it is washed in 95% alcohol and is used for 55 scraping only. Aftereach scraping the scalpel is washed with 95% alcohol, the alcohol isremoved by sterile cotton, and the scalpel is then put back into the 95%phenol solution.

The instrument for removing the serum consists of a shovel-likeflattened scoop made of platinum, about two inches long, and connectedto a suitable handle. This instrument is sterilized by holding it in asuitable flame and the scoop is thrust upon the scarified surface andpressed forcibly to express the serum or lymph which is then taken up bythe scoop and directly inoculated into the culture medium.

After each use thereof, the last mentioned instrument, together with thesteel handle thereof, is sterilized by holding it in a suitable flame.

The above mentioned procedure is important in securing a substantiallypure yield of the micro-organism which produces this disease, and I havefound that I can secure a yield of over 90%.

In addition to securing scales and serum as above mentioned, bloodcultures and biopsies can also be taken. For this purpose of takingblood, a suitable surface of the forearm is washed with soap and water,it is then washed with 70% alcohol and then with 95% alcohol, and it isthen washed with ether to thoroughly remove the alcohol and any fat. Thesurface is then washed by a suitable disinfectant, such as a solution ofmercurochrome in alcohol. This solution may be prepared by saturating95% alcohol with mercurochrome and using one part of this solution to1000 parts by volume of 50% alcohol. The surface is then covered with asterile dressing for half an hour. After this, the vein is located inthe cubital fossa and with a sterile needle and syringe 15 cc. of bloodis obtained which is introduced into flasks of maltose agar and kept inincubator until white colonies are seen through the glass (sometimes aslong as 6 weeks).

As previously mentioned, the reason for carefully sterilizing the tissueprior to taking materials for cultures is to reduce the liability ofsurface contamination and to secure the causative agent of the diseasein as pure a condition as possible.

The blood culture obtained is preferably inoculated directly into anErlenmeyer flask containing a sugar agar-slant. This agar-slant has asurface of substantial area which is inclined to the axis of the flask,so that the micro-organism reproduces freely upon a large exposedsurface of the agar and can be detected by means of such surfacereproduction, as will be later more fully explained. This micro-organismcan be allowed to attain the necessary growth without opening the flask,which lessens the danger of contamination, and it can also be detectedby means of its surface growth without opening the flask. Ordinarily inpropagating blood cultures in a flask it is necessary to open the flaskat fairly frequent intervals to make repeated tests. Of course, eachopening of the flask increases the danger of contamination, which isentirely obliviated by the procedure above specified.

In order to observe whether or not the filamentous causativemicro-organisms have been stcured, first hanging-drop monospore culturesare studied and thus the different phases of growth and reproduction inwhich they have been secured are observed. Then permanent smears may beobserved microscopically. All smear preparations directly made from thelesions, and also including those made from the cultures to observe the'reproduction of the micro-organisms, are dried in the air, fixed byflame and by a mixture containing equal parts of ether and 95% alcohol.The use of the alcohol-ether mixture is to remove fats. The smears arethen stained by any suitable method and I prefer to use Giemsas method.The Giemsa stain, diluted 60 drops to 100 cc. distilled water, can bedirectly applied on the smear from 15 to 50 minutes or can be utilizedovernight. The Giemsa stain has heretofore been used for staining tissuespecimens, in which case the stain was washed out by means of acetoneand xylol. But in my method for smears the stain is merely washed outwith water.

In the same manner smears prepared by impression of the irritatedsurface of the lesion and from fragments cut from the biopsies obtainedfrom the psoriatic lesions were stained. It is worth noting that thefragments of the psoriatic biopsies first washed in 60% alcohol for anhour and then incubated at 375 C. gave both a positive growth andmicroscopic finding in the smears. That is, the characteristicappearance of the filamentous micro-organism, which I believe to be thecausative agent of the disease, was noted culturally and microscopicallyfrom both the scrapings as well as the fragments of the biopsies in theannexed drawing including Nos. 1, 2, 3, 4, 5, 6, 7, 8 and 9. In thebiopsies similar Figures to l, 5 and 6 as shown in said drawing wereobserved.

When this filamentous micro-organism reproduces under favorableconditions and proper culture media with a pH 6.4 at 37.5 C. it forms awhite surface layer upon the culture media, which can be readilyobserved by the naked eye after suflicient time has elapsed. Of course,the reproductions have variations dependent upon the patient from whomthe specimens have been taken and upon other conditions, some of whichhave not yet been determined. Ordinarily under favorable conditions thesaid white surface layer which consists mostly of spores can be detectedafter a lapse of four days and even less as may be noted in thephysiological study of the microorgamsm.

Physiological study of the Actinomyces psoriaticum Generally thismicro-organism can be propagated in various culture media of differentmodified sugars glycerine, either solid agar or bouillon, milk, potatoand various synthetic, serum, whole blood and albumin or egg at varioustemperatures according to the preference of the culture age andbehavior, but ranging from below 20 C. to above 40 C. for several days.

For descriptive study it is preferable to use body temperature (375 C.)and specifying the appearance of the individual colony or the entireculture as it may be necessary within every few hours if possible on thevarious culture media. It was thus observed that when the micro-organismis grown of glycerine agar or bouillon the colony appears somewhatlarger. Even when subcultures are made from glycerine into other sugarmedia the usual size is increased. In general, under favorableconditions, the colony before fructification, thatis, before the white,porcelain, powdery surface takes place, appears round, elevated, uniformin size, gradually assuming a dark green color, penetrating deep intothe substratum of the culture medium in from 24 to 48 hours and measuresfrom 3 to 5; microns in diameter, but as soon as fructificatio'n occursthe size of the colony inwhich the micro-organism has been grown andcreases; unless the micro-organism should be in the pleomorphic stagewhen the colony may become larger without assuming the white surface ofspores. More specifically, unless the colony is in a pleomorphic stage,it is usually smaller before the production of the white layer ofspores; and larger after such fructification takes place. This whitelayer consists purely of the spores and the layer under the spores arethe filaments which penetrate deep into the substratum of the culturemedium. Thus the dark green color of the colony takes place beforefructlfication, that is before the production of the individual spores.

The growth may also difier on the diiferent culture where the degree ofthe tyrosinase reaction may also vary in the depth and concentration ofthe dark brown to black color, indicating the moderate, marked orvigorous growth of the micro-organism, Lehman, K. 13.: Uber das Vormmenvon Oxydations fermenten bei Bacterien und Sanum Heren Pflanzen, Arch,Hygiene, 6'? 99 113.

Stab culture in sugar agar or in aelatine Most of the strains have grownon the surface as well as in the depth of the medium. The growth appearsas a dense beaded dark green streak if before fructification in thefirst 24 or 36 hours at 3'1.5 C. or the streak becomes white if thisperiod of incubation exceeds 36 hours. These colonies may also appear assmall nodules either on the line of inoculation or on the surface wherethey are likely to be more dense somewhat spreading into the substratumwith a greenish base. The tyrosinase reaction usually takes place withinsix or ten days of growth varying with the virulence or age of themicro-organism.

Bouillon culture either with the addition of carious sugars or with theaddition of various chemicals-The growth also varies with time 'ofincubation as above, but in general the growth appears either at thebegining after 24-38 hours of incubation at 37.5 as solid single ormasses of bodies throughout the medium; some float on the surface,others cling to the sides of the tube and still others may sink to thebottom, but the bouillon is always clear unless contamination takesplace. I

Litmus milk-Litmus was peptinized within six days at 375 C. Thereactions appeared to be irregularsome alkaline and others acid.

Egg albumin.-The growth on egg albumin appeared to be very fine,markedly elevated round colonies, which penetrated the substratum of theculture medium. Hydrolysis occasionally occurred as well as thetyrocinase reaction.

Czopaclcs synthetic culture medium-This medium occasionally showed veryfine shades of chromogenic difl'erences, but the surface colonies alsoappeared very fine as on the egg albumin, which are not suitable forgeneral description. Solid culture of carbohydrates were thereforenecessary for the study of general description.

Details of the growth observed on glucose agar or double or triple sugarand glycerine are similar to those described under solid and bouillonculture media.

More details will be described under the morphological study as well asunder the treatment by the inoculation of the living micro-organismobtained from the surface growth of solid media and inoculated into thescarified sound skin of a the patient and also under the preparation ofspecific psoriasis vaccine which can be prepared either from solid orbouillon culture media upon obtained inpure culture after it has beensubcultured a number of times.

The following studies are given as an explanation and description of thespecific organism causing psoriasis which is found and the methods bywhich it may be applied to treat the disease.

Morphology of Actinomyces psoriaticum Morphologically the "Actinomyces'psoriaticum has points of resemblance to Actinomyces VIII, described byDrechsler (Morphology of the Genus Actinomyces, The Chemical Gazette,Nos. 1 and 2, January and February, 1919) and culturally resemblesActinomyces V Stamm 39, de-' scribed by Lieske (Morphologie und Biologiedes Stralenpilze. Pages 2'79 and 293, Heidelburg, 0ctober, 1920,Botanische Institut) and as noted in the drawing. The characteristicswhich retain the identity of Actinomucespsoriaticum will be pointed outbelow:

Although other Actinomuces of different sources have been studied alongwith Actinomyees psoriaticum only the latter showed characteristics thatmay be essential in order to differentiate it from the other Actinomycesof other sources than psoriasis.

Five strains of the "Actinomyces psoriaticum monospore cultures havebeen studied in the hanging-drop-wet-chamber preparation. Under suitableconditions in glucose agar the microorganism from all the five strainsgrew essentially in the same manner in the hanging-dropcultures. If thewater in the bottom was sufficient to prevent the hanging-drop fromdrying, and the cultures were kept continuously in the incubator,thereoccurred a branching filamentous growth in two days. Gradually,more or less, erect fructification developed along the distal portion;the fertile hyphae at times appeared to be attached to the axialfilaments in a diffuse system. In the course of about a week or more theswellings in the axial filaments at the base of the sporagenous branchesindicate the site of origin, and the germination of some separatedslnglespores or clusters of spores is also one of the characteristics ofidentification. Besides those characteristics of the grouping of thespores, the size and shape of the spores are also points of differentialclassification:-The spores are ellipsoidal showing a definite centralbody which specifies to call the spores uninucleated, measuring 0.5 to1.3 microns in diameter. If about 1% phosphate is added to the culturemedia, there develop-close sinistral spirals consisting of 2-4 turns,measuring 1.5 to 2.5 microns in diameter; also it carefully studied,characteristic cork-screw bodies are observed which consist of 2-4 loopsand 1-3 knobs, as shown at 6 in the drawing.

It is thus noted that the limited and definite grouping. size andshape-oi. the spores; the limited number of spirals (2-4) the limitednumber of loops (2-4) as well as the limited number of knobs (l-3) arethe characteristic points of differential classification and thereforethese points retain the identity of the Actinomyccs psoriaticum.

The Actinomyces described by Drechsler show a greater number of thespirals, cork-screw bodies and knobs than the Actinomyces psoriaticumwhile the Actinomyces described by Lieske did not show any of thesecharacteristics in his report.

The parts illustrated in the drawing are identified as follows:

1. A single spore.

2. Filaments of a germinated spore.

3. A sporogenous branch with terminal spirals.

4. A germinating spore with germ-tube.

5. Aerial mycilium hooked over with the spiral hyphae of another branch.

6. Spirals and cork-screw bodies showing definite loops in groups ofthree. The same number (three) of loops was observed in another cultureand in a section of a biopsy taken from-a psoriasis lesion which alsoshowed same number of three loops.

'7. Germinating spores with fructification developed in successivesequence with chains and single spores with germ-tube.

8. Fertile aerial mycilium showing spore indieating origin ofsporogenous short branch of mycilium consisting of immature spores.

9. Aerial germinating spore with two germtubes, originating from myciliaincluding immature spores.

Therapeutic properties of "Actinomg/ces psoriaticum detected During theexperiment in carrying out Koch's postulate on psoriatics andnon-psoriatics as well as animals (including monkeys and rabbits) bydirect inoculation of the living micro-organisms obtained from thesurface growth of solid culture media at different stages of theActinomyces psoriaticum, it was noted that the pre-existing lesions inthe psoriatics either completely or partially cleared up after abouteight series of inoculations into the scarified skin were made duringtwo months.

With further persistent effort, it was noted that these inoculationsmade into scarified skin in quiescent psoriasis cases, the active stagetook place after six series of inoculations with the livingmicro-organism into the scarified skin. The first series was startedwith three scariflcation inoculations and gradually increased with oneadditional scariflcation at a time, until ten such inoculations weremade at the last series. This usually gave rise to an active stage atwhich time the clearing of the original lesions were observed.

Hence I believe that this micro-organism possessed therapeuticproperties and a vaccine therapy as well as thescarification-inoculation treatment was tried with successful results.

These inoculations were made purely from the surface growth of themicro-organism from solid media which was readily removed from theculture medium. These spores do not have a shape which distinguishesthem from the genus Actinomyces.

Directly underneath this easily removed surface layer, a filamentousstructure is found which has penetrated and is closely associated withthe culture medium, so that it is not possible to remove thisfilamentous structure without taking along some of the culture mediumsimultaneously.

The surface layer of the culture, which comprises both the filamentousmicro-organism and the living spores and filaments, can be used directlyby scarification and then by gently rubbing in the culture medium sothat it is absorbed by the blood of the patient. This medium should beapplied by means of the platinum scoop previously mentioned.

However, as this requires very tedious and careful manipulation,although it is therapeutically effective, I prefer to use the vaccinelater mentioned, although direct application of the living culture isoften of great value in quickly producing the active stage of thedisease in a patient, so that the vaccine is then more effective. I havediscovered that when psoriasis lesions are in the active stage thevaccine treatment is more effective. I have further discovered that thedirect inoculation of the living organism isolated from psoriasis intoquiescent psoriatics produces more active psoriasis lesions on soundskin. This more active form lends itself to the vaccine treatment morereadily than the quiescent form.

In order to prepare the vaccine, the subcultures are prepared upon theslant-agarsurface from the initial growth previously mentioned(preferably in an Erlenmeyer flask) and they are allowed to germinatefor only about two days at 37 C. and before fructification has takenplace with the formation of the white surface layer previouslymentioned.

The surface of the agar culture now contains the colonies in filamentousand germinating form and these cling so firmly to the said surface ofthe agar that the surface portion of the agar must be cut out in orderto remove the germinating colonies.

The culture thus secured is collected in previously sterilized glassbottles which contain sterilized glass beads and a normal salinesolution (NaCi0.85%) intermixed with a .3% phenol solution.

The cultures from several patients are preferably intermixed in one ofthese bottles so as to produce a polyvalent vaccine. I wish to state atthis point that even following all the precautions above mentioned, itis not always possible to secure a culture from a patient, and that theease of obtaining cultures from various patients differs with individualcharacteristics.

A vaccine can also be produced from the culture containing both thespores and the filaments of the micro-organisms, but is less reliable,and

hence I prefer to use the first mentioned vaccine. I

The several strains of culture are now vigorously shaken, preferably bymachine, from two to four days and even more, until the said suspensionis homogenous.

The several strains of the culture are now vigorously shaken, by specialmachine, from twentyfour to forty-eight hours and even longer, until thesaid suspension is homogenous and is free from clots, so that a dropthereof, free from clots, can be readily taken up by the finesthypodermic needle.

The consistency of the vaccine is now determined by comparing theviscosity of a standard vaccine, such as the Staphylococcus vaccine,having 10,000 per cc.

The suspension is now made sterile either by heating it at a temperatureof 60 C. for three hours, followed by an addition of 0.5% phenol; or thephenol and heat is not used at all, but instead the saline (0.85%) usedfor the suspension is first treated with 1:5000 merthiolate and thenwhen the micro-organism is suspended and shaken for a sufficient periodof time (24-48 hours) the suspension is incubated for three days at 375C. and then tested for sterility. A number of other chemicals have beensuccessfully tried in effecting sterility without the heating of thesuspension. Among these chemicals were included phenol and creosote,etc., but it was found that the merthiolate is most efiicient as thereit causes no breaking down of the constituents into protein. Andfurthermore, after sterility is effected with the 1:5000 dilution of the"merthiopared either from the propagated micro-organism on the variousagar-agar culture media, or in various modified bouillons, including allthe combinations of sugars, blood, blood serum and glycerine or thesynthetic media which need not have an exposed surface. At the end of 38or 48 hours appear the characteristic round, dense beaded streaks (orlines of small nodules) along the line of inoculation. In bouillon thesecolonies may float on the surface or cling to the sides of the tube. Itthe growth is prolonged the colonies become white masses or individualgroups of bodies or both on the surface and throughout the medium. Forthe purpose of preparing the suspensions or the extracts it is necessaryto remove the growth of the micro-organism either from the solid culturemedia or from the bouillon before white growth or the individual sporesare formed which can be seen by microscopic examination not before the36 hours of growth. If the growth is thus prepared in bouillon culturemedia it should be removed either by syphoning oil the bouillon or byfiltering it oil, and retaining the micro-organism on the filter, whichis then used either for the suspension or for extracts, as well as formicroscopic characteristics in order to detect and eliminate the spores.

It is absolutely necessary substantially either to inhibit the formationof the spores, which is preferable, or to eliminate spores in theproduction of the vaccine; because the presence of spores has been foundto produce certain undesirable incidental effects, such as theproduction of abscesses or the like.

Thus the extract vaccine which is an extract and a filtrate of themicro-organism "Actinomyces psoriaticum is preferable for thetherapeutic purpose.

Method of preparing psoriasis extract I have also succeeded in preparingan extract of psoriasis vaccine. This extract is obtained from the"Actinomyces psoriaticum. The above suspension prepared for the specificpsoriasis vaccine" is also applied for the preparation of the extract,but without shaking. Thus, the suspension of the spore-free filamentouscolonies obtained from the growth of the "Actinomuces psoriaticum in thevehicle-0.85% saline containing 2.5% phenol-is kept in the incubator at375 C. for 48 hours, then tested for sterility and without shaking issubjected to extraction with sterile 0.85% saline at intervals of twoweeks, in a course of eight weeks, and then subjected to the Berkefeldfilter. This extract is thus diluted to the extent where it reaches toretain only 0.5% phenol, but further dilutions may be necessary whenapplied for testing.

This extract is eificient either for treatment or for testing,-subcutaneously and intradermally, respectively. The purpose of testingwith this extract is to determine the degree of susceptibility or thedegree of the immunity established during the course of treatment eitherwith the "specific psoriasis vaccine or with the extract, respectively,or both used alternatively.

I claim:

1. In preparing a polyvalent vaccine for the treatment of psoriasis thesteps which consist in treating papules of said disease with asterilizing medium, securing from said papules a. filamentousmicro-organism herein referred to as Actinomuces psoriaticum, saidmicro-organism being capable of reproducing in culture media having a pHof 6.4 at temperatures between 20 and 40 C. and forming round dark greencolonies measuring from 3 to 5 microns in diameter, said coloniesincreasing in size and acquiring a white powdery surface whenfructification occurs, said micro-organisms having ellipsoidal sporesmeasuring from ,5 to'1.3 microns in diameter, said micro-organisms alsocontaining bodies having close sinistral spirals of 2-4 turns andcorkscrewshaped bodies having 2-4 loops and 1-3 knobs when 1% phosphateis added to the medium containing said micro-organism, propagating saidmicro-organism in a culture medium, preparing sub-cultures therefrom,separating the sporefree filamentous germinating form of the saidmicro-organism from the subculture medium, intermixing said spore-freefilamentous germinating form of the said micro-organism with otherstrains of the same form of said micro-organism from other sources,suspending these mixed strains in a liquid vehicle, homogenizing thesus-" pension, and sterilizing the said suspension.

2. In preparing a vaccine for treatment of psoriasis, the steps whichconsist in treating infected papules of said disease with a sterilizingmedium, securing from said papules a filamentous micro-organism hereinreferred to as Actinomj ces psoriaticum, said micro-organism beingcapable of reproducing in culture media having a pH of 6.4 attemperatures between 20 and 40 C. and forming round dark green coloniesmeasuring from 3 to 5 microns in diameter, said colonies increasing insize and acquiring a white powdery surface when fructification occurs,said micro-organisms having ellipsoidal spores measuring from .5 to 1.3-microns in diameter, said micro-organisms also containing bodies havingclose sinistral spirals of 2-4 turns and corkscrew shaped bodies having2-4 loops and 1-3 knobs when 1% phosphate is added to the mediumcontaining said micro-organism, propagating said micro-organism in aculture medium, preparing sub-cultures therefrom, separating thesporefree filamentous germinating form of the said micro-organism fromthe sub-culture medium, suspending this micro-organism in a liquidvehicle, homogenizing the suspension and sterilizing the saidsuspension.

3. In preparing a polyvalent vaccine for the treatment of psoriasis, thesteps which consists in treating infected papules of said disease with asterilizing medium, securing from biopsies of said papules offilamentous microorganism herein referred to as Actinomyces psoriaticum,said microorganism being capable of reproducing in culture media havinga pH of 6.4 at temperatures between 20 and 40 C. and forming round darkgreen loops and 1-3 knobs when 1% phosphate is added to the mediumcontaining said micro-organism. propagating said micro-organism in aculture medium, preparing subcultures therefrom, separating thespore-free filamentous germinating form of the said micro-organism fromthe subculture medium, intermlxing said spore-free filamentousgerminating form of the said mieroorganism with other strains of thesame form of said micro-organism from other sources, suspending thesemixed strains in a liquid vehicle, homogenizing the suspension, andsterilizing the said suspension.

4. In preparing a polyvalent vaccine for the treatment of psoriasis, thesteps which consist in treating infected papules of said disease with asterilizing medium, securing from said papules a filamentousmicro-organism herein referred to as Actz'nomyces psoriaticum, saidmicro-organism being capable of reproducing in culture media having a pHof 6.4 at temperatures between 20 and 40 C. and forming round dark greencolonies measuring from 3 to 5 microns in diameter, said coloniesincreasing in size and acquiring a white powdery surface whenfructification occurs, said micro-organisms having ellipsoidal sporesmeasuring from .5 to 1.3 microns in diameter, said micro-organisms alsocontaining bodies having close sinistral spirals of 2-4 turns andcorkscrew shaped bodies having 2-4 loops and 1-3 knobs when 1% phosphateis added to the medium containing said micro-organism, progagating saidmicro-organism in a culture medium, preparing sub-cultures therefrom,separating the spore-free filamentous germinating form of the saidmicro-organism from the sub-culture medium, intermixing said spore-freefilamentous germinating form of said micro-organism with other strainsof the same form of said microorganism, suspending these mixed strainsin a liquid vehicle, adding merthiolate to said suspension, incubatingto kill said micro-organism and homogenizing said suspension.

5. In preparing an agent for the treatment of psoriasis those stepswhich consist in treating infected papules of said disease with asterilizing medium, securing from said papules a filamentousmicro-organism herein referred to as Actinomyces psoriaticum, saidmicro-organism being capable of reproducing in culture media having a pHof 6.4 at temperatures between 20 and 40 C. and forming round dark greencolonies measuring from 3 to 5 microns in diameter, said coloniesincreasing in size and acquiring a white powdery surface whenfructification occurs, said microorganisms having ellipsoidal sporesmeasuring from .5 to 1.3 microns in diameter, said microorganisms alsocontaining bodies having close sinistral spirals of 2-4 turns andcorkscrew shaped bodies having 2-4 loops and 1-3 knobs when 1% phosphateis added to the medium containing said micro-organism, propagating saidmicroorganism in a culture medium having an exposed surface, separatingfrom said culture medium the spores found adjacent to said exposedsurface and separating from said culture medium the exposed surfaceportions thereof containing spores and filamentous micro-organism.

6. In preparing an agent for the treatment of psoriasis those stepswhich consist in treating infected papules of said disease with asterilizing medium, securing from the various levels of said papules afilamentous micro-organism herein referred to as Actinomycespsoriaticum, said micro-organism being capable of reproducing in culturemedia having a pH of 6.4 at temperatures between 20 and 40 C. andforming round dark green colonies measuring from 3 to 5 microns indiameter, said colonies increasing in size and acquiring a white powderysurface when fructification occurs, said micro-organisms havingellipsoidal spores measuring from .5 to 1.3 microns in diameter, saidmicro-organisms also containing bodies having close sinistral spirals of2-4 turns and corkscrew shaped bodies having 2-4 loops and 1-3 knobswhen 1% phosphate is added to the medium containing said micro-organism,progagating said micro-organism in a culture medium having an exposedsurface, separating from said culture medium the spores found adja centto said exposed surface and separating from said culture medium theexposed surface portions thereof containing spores and filamentousmicroorganism.

7. In preparing an agent for the treatment of psoriasis those stepswhich consist in treating infected papules of said disease with asterilizing medium, securing from the biopsies of said papules afilamentous micro-organism herein referred to as Actinomycespsoriatz'cum, said micro-organism being capable of reproducing inculture media having a pH of 6.4 at temperatures between 20 and 40 C.and forming round dark green colonies measuring from 3 to 5 microns indiameter, said colonies increasing in size and acquiring a white powderysurface when fructification occurs, said micro-organisms havingellipsoidal spores measuring from .5 to 1.3 microns in diameter, saidmicro-organisms also containing bodies having close sinistral spirals of2-4 turns and corkscrew shaped bodies having 2-4 loops and 1-3 knobswhen 1% phosphate is added to the medium containing said micro-organism,propagating said micro-organism in a culture medium having an exposedsurface, separating from said culture medium the spores found adjacentto said exposed surface and separating from said culture medium theexposed surface portions thereof containing spores and filamentousmicro-organism.

8. A sterile vaccine for the treatment of psoriasis comprising asuspension of a sport-free substantially filamentous micro-organismherein referred to as Actinomyces psoriaticum, said micro-organism beingcapable of reproducing in culture media having a pH of 6.4 attemperatures between 20 and 40 C. and forming round dark green coloniesmeasuring from 3 to 5 microns in diameter, said colonies increasing insize and acquiring a white powdery surface when fructification occurs,said micro-organisms having ellipsoidal spores measuring from .5 to 1.3microns in diameter, said micro-organisms also containing bodies havingclose sinistral spirals of 2-4 turns and corkscrew shaped bodies having2-4 loops and 1-3 knobs when 1% phosphate is added to the mediumcontaining said microorganism, said suspensions being prepared accordingto the steps consisting of treating papules of said disease with asterilizing medium, securing said micro-organism from said papules,propagating said micro-organism, preparing a sub-culture therefrom,separating the spore free filamentous germinating form of the saidmicroorganism from the subculture medium, suspending said micro-organismobtained from the subculture in a liquid vehicle, homogenizing thesuspension and sterilizing the said suspension.

9. A sterile polyvalent vaccine for the treatment of psoriasiscomprising a suspension of a spore-free substantially filamentousmicro-organism herein referred to as Actinomyces psorzaticum, saidmicro-organism being capable of reproducing in culture media having a pHof 6.4 at temperatures between 20 and C. and forming round dark greencolonies measuring from 3 to 5 microns in diameter, said coloniesincreasing in size and acquiring a white powdery surface whenfructification occurs, said microorganisms having ellipsoidal sporesmeasuring from .5 to 1.3 microns in diameter, said microorganisms alsocontaining bodies having close sinistral spirals of 2-4 turns andcorkscrew shaped bodies having 2-4 loops and l-3 knobs when 1% phosphateis added to the medium containing said micro-organisms, said suspensionbeing prepared according to the steps consisting of treating papules ofsaid disease with a sterilizing medium, securing said micro-organismfrom said papules, propagating said micro-organism, preparing asub-culture therefrom, separating the spore-free filamentous germinatingform of the said micro-organism from the sub-culture medium, intermixingsaid micro-organism obtained from the sub-culture with other strainsobtained in the same manner, suspending these mixed strains in a liquidvehicle, homogenizing the suspension and sterilizing the saidsuspension.

10. A sterile vaccine for the treatment of psoriasis comprising apolyvalent extract and filtrate of a spore-free substantiallyfilamentous micro-organism herein referred to as Actinomycespsoriaticum, said micro-organism being capable of reproducing in culturemedia having a pH of 6.4 at temperatures between 20 and 40 C. andforming round dark green colonies measuring from 3 to 5 microns indiameter, said colonies increasing in size and acquiring a white powderysurface when fructification occurs, said micro-organisms havingellipsoidal spores measuring from .5 to 1.3 microns in diameter, saidmicro-organisms also containing bodies having close sinistral spirals of2-4 turns and corkscrew shaped bodies having 2-4 loops and 1-3 knobswhen 1% phosphate is added to the medium containing said micro-organism,said extract being prepared according to vthe steps consisting oftreating papules of said disease with a sterilizing medium, securingsaid micro-organism from said papules, propagating said micro-organism,preparing a sub-culture therefrom, separating the spore-free filamentousgerminating form of the said micro-organism from the sub-culture medium,suspending said micro-organism obtained from the sub-culture in a liquidvehicle, sterilizing the said suspension without shaking it, andpreparing an extract and filtrate therefrom.

11. In preparing a vaccine for the treatment of txammel psoriasis, thesteps which consist in sterilizing a portion of sound skin, obtaining asample of the psoriatic patients blood through the sterile portion ofthe said skin, securing from said blood a filamentous micro-organismherein referred to as Actinomyces psoriaticum, said micro-organism beingcapable of reproducing in culture media having a pH of 6.4 attemperatures between 20 and 40 C. and forming round dark green coloniesmeasuring from 3 to 5 microns in diameter, said colonies increasing insize and acquiring a white powdery surface when fructification occurs,said micro-organisms having ellipsoidal spores measuring from .5 to 1.3microns in diameter, said micro-organisms also containing bodies havingclose sinistral spirals of 2-4 turns and corkscrew shaped bodies having2-4 loops and 1-3 knobs when 1% phosphateis added to the mediumcontaining said micro-organism, propagating said micro-organism in aculture medium, preparing sub-cultures therefrom, separating thespore-free filamentous germinating form of the said micro-organism fromthe sub-culture medium, intermixing said spore-free filamentousgerminating form of the said micro-organism from other sources,suspending these mixed strains in a liquid vehicle, homogenizing thesuspension, and sterilizing the said suspension.

12. A sterile vaccine for the treatment of psoriasis comprising asuspension of the sporefree substantially filamentous micro-organismherein referred to as Aciinomyces psoriaticum, said micro-organism beingcapable of reproducing in culture media having a pH of 6.4 attemperatures between 20 and 40 C. and forming round dark green coloniesmeasuring from 3 to 5 microns in diameter, said colonies increasing insize and acquiring a white powdery surface when fructification occurs,said micro-organisms having ellipsoidal spores measuring from .5 to.1.3microns in diameter, said micro-organisms also containing bodies havingclose sinistral spirals of 2-4 turns and corkscrew shaped bodies having2-4 loops and 1-3 knobs when 1% phosphate is added to the mediumcontaining said microorganism, said suspension being prepared accordingto the steps consisting in sterilizing a portion of the sound skin,obtaining a sample of the psoriatic patients blood through the saidskin, propagating the said micro-organism contained in said blood in aculture medium, preparing sub-cultures therefrom, separating thesporefree filamentous germinating form of the said micro-organism fromthe sub-culture medium, suspending said micro-organism obtained from thesub-culture in a liquid vehicle, homogenizing the suspension andsterilizing the said suspension.

MARY A. MARCUS.

